BAC Library Production for Comparative Genetics

比较遗传学的 BAC 文库生产

• 批准号: 6805895
• 负责人: Pieter J. de Jong
• 金额: $ 15万
• 依托单位: CHILDREN'S HOSPITAL & RES CTR AT OAKLAND
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-22 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6805895
• 关键词: artificial chromosomes; biotechnology; comparative genomic hybridization; cooperative study; eukaryote; functional /structural genomics; genetic library; high throughput technology; microarray technology; molecular cloning; nucleic acid sequence; restriction fragment length polymorphism; technology /technique development

The goal is to prepare BAC libraries for many species, of high quality, tailored to the scientific community's interest in comparative genomics and functional applications. The work will be done by two collaborating teams, one lead by Dr. Cheng at the Lawrence Berkeley National Laboratory (LBNL) and the other one at the Children's Hospital Oakland Research Institute, co-directed by Drs. Osoegawa & deJong. The application has four components relevant to preparing Bacterial Artificial Chromosomes (BACs): 1) BAC Library preparation, 2) Clone Arraying & Library Duplication, 3) Library Characterization, and 4) Research & Development to improve the overall process efficiency and the BAC library quality. During the first year, the consortium will generate and characterize twelve animal BAC libraries with ten-fold genome redundancy. Presuming an increasing need for additional GBACs and improved production efficiency, 17 and 22 additional libraries will be prepared and characterized in years 2 and 3 respectively. The libraries, based on current BAC cloning technology, will complement and extend the small repertoire of BAC clone collections now available for comparative genome analysis. The new clone collections will be analyzed by a standardized set of tests, including screening with a set of economic markers, limited BAC-end sequencing, BAC fingerprinting, BAC stability analysis and insert size determination. Of particular importance are quality tests to ensure low BAC stability analysis and insert size determination. Of particular importance are quality tests to ensure low levels of clonal cross contamination. Once the libraries pass the quality controls, the new resources will be made available in a format consistent with major applications. To improve the overall process efficiency, a set of Standard Operating Procedures (SOP's) will be developed to move BAC cloning away from an art form towards a routine high-throughput procedures. It is expected that the new BAC resources will contributed towards a routine-high throughput procedure. It is expected that the new BAC resources will contribute to better understanding of gene function and evolution through comparison of genes in a spectrum of animal species. The BAC clones will also become tools to create animal models for human diseases. Widespread dissemination of the clones is an unfunded but essential component of this proposal. Library information will be disseminated by publication and through our home page (http://www.chori.org/bacpac).

目标是为许多物种准备高质量的BAC文库，以满足科学界对比较基因组学和功能应用的兴趣。这项工作将由两个合作团队完成，一个由劳伦斯伯克利国家实验室（LBNL）的Cheng博士领导，另一个由Osoegawa博士和deJong博士共同指导的儿童医院奥克兰研究所领导。该应用程序有四个与制备细菌人工染色体（BAC）相关的组件：1）BAC文库制备，2）克隆阵列和文库复制，3）文库表征和4）研究与开发，以提高整体工艺效率和BAC文库质量。在第一年，该联盟将产生和表征12个具有10倍基因组冗余的动物BAC文库。假定对额外GBAC的需求不断增加，生产效率也不断提高，将在第2年和第3年分别制备和表征17个和22个额外的文库。这些基于当前BAC克隆技术的图书馆将补充和扩展目前可用于比较基因组分析的小规模BAC克隆库。将通过一套标准化的测试对新的克隆集合进行分析，包括用一套经济标记进行筛选、有限的BAC末端测序、BAC指纹分析、BAC稳定性分析和插入片段大小确定。特别重要的是质量测试，以确保低BAC稳定性分析和插入尺寸确定。特别重要的是质量测试，以确保低水平的克隆交叉污染。一旦图书馆通过质量控制，新资源将以与主要应用程序一致的格式提供。为了提高整体工艺效率，将开发一套标准操作规程（SOP），以将BAC克隆从艺术形式转变为常规高通量程序。预计新的BAC资源将有助于常规高通量程序。预计新的BAC资源将有助于通过比较一系列动物物种中的基因来更好地理解基因功能和进化。BAC克隆还将成为创建人类疾病动物模型的工具。克隆人的广泛传播是这项提议的一个没有资金支持但必不可少的组成部分。图书馆信息将通过出版物和我们的主页（http：//www.choria.org/bacpac）传播。